The Study Of Intestinal Immune And Overactive Immune Mechanism Of Kidney-Nourishing, Spleen-Strengthening And Detoxification Therapy For SIV/AIDS

ObjectiveIn this paper, kidney-nourishing, spleen-strengthening and detoxification therapy was enforced on Chinese rhesus AIDS model to investigate in the following two aspects:1. Explore the mechanism of TCM intervention in gut lymphoid homing. Find out the strategy of TCM intervention of treating AIDS in the perspective of intestinal mucosal immunity.2. Investigate the mechanism of TCM intervention of overactive immune system, and the balance between immune activation and immune tolerance.MethodsWhen the monkeys used in the experiment were successfully modeled, we tested the monkey’s body weight, blood routine, blood biochemistry, viral load, T lymphocyte subsets as well as by radioimmunoassay of serum T3(triiodo thyronine), T4(thyroxine), COR (cortisol) and β2-MG (serum β2microglobulin) routinely, which proved that the monkeys have had the characteristics of spleen and kidney deficiency.Specific contents of the experimental monkeys were as follows:1. The study on the activation of gut-associated lymphoid tissue and the expression of intestinal lymphoid homing receptor on the gut-associated lymphoid tissue:a total of16monkeys were randomly divided into treatment group and model control group (n=8). The experiment builds on the previous study results, to depth investigate kidney-nourishing, spleen-strengthening and detoxification interventions in intestinal lymphatic homing mechanism and overactive immune system. Immunohistochemical techniques were used to detect the expression of the gut homing related-receptor α4β7, CCR9, αEβ7, CD62L and intestinal lymphoid tissue activation molecules CD69on the ileal lymphoid tissue.Using ELISA technique to detect the changes of CD62L plasma concentrations.2. The study on the activation of peripheral blood lymphocytes and the expression of gut homing receptor on peripheral blood lymphocytes:total15monkeys are infected through intravenous SIVmac239and successfully modeled. They were randomly divided into three groups:low-dose group, high dose group and normal saline control group (n=5). This paper focus on①investigate the activation of peripheral blood T lymphocytes, the expression of gut associated lymphoid homing receptor αEβ7、CCR9and plasma concentrations changes of CD62L in HIV acute phase monkey;②depth study of kidney-nourishing, spleen-strengthening and detoxification intervention the AIDS monkeys in immune activation conditions, CD62L plasma concentrations changes, intestinal inflammation-related factors LPS activity conditions. Use flow cytometry techique to detect the expression of HLA-DR molecules on T lymphocyte; use enzyme-linked immunosorbent assay (ELISA) techique to detect CD62L plasma concentrations and serum concentrations of LPS; The detection time were before infection, before administration (infection11weeks), administered2months and withdrawal2months.Results1. The expression of CD69molecule in intestinal lymphoid tissue after kidney-nourishing, spleen-strengthening and detoxification therapymodel group of100days compared with the baseline point of model group, the expression of CD69molecule on ileal tissue was significantly higher (P=0.000); medication50days compared with the baseline point of model group, the expression of CD69molecule decreased (P=0.032); medication100days compared with model group of100days, CD69molecule expression was significantly decreased (P=0.01); treatment100days compared with the treatment of50days, CD69molecule expression was significantly higher (P=0.01), but significantly lower than the model group expression levels at the same time point (P=0.01).2. The expression of gut lymphoid homing receptor after kidney-nourishing, spleen-strengthening and detoxification therapy2.1The gut homing receptor CCR9molecule expression of ileal lymphoid tissue Model group of100days compared with the baseline point of model group, intestinal lymphoid homing receptor CCR9expression of ileal tissue was significantly higher (P=0.005); medication50days compared with the baseline point of model group, CCR9molecule expression was significantly decreased (P＝0.003); medication100days compared with model group of100days, CCR9molecule expression decreased (P=0.040); treatment100days compared with the treatment of50days, CCR9molecule expression was significantly higher (P=0.000), but significantly lower than the model group at the same time point expression (P=0.000)2.2ileal lymphoid tissue gut homing receptor molecule αEβ7expressionModel group100days compared with the baseline point of model group, ileal lymphoid tissue gut homing receptor αEβ7expression was significantly increased (P=0.003); medication50days compared with the baseline point of model group, αEβ7expression was significantly decreased (P=0.008); medication100days compared with the model group100days, αEβ7expression decreased (P=0.012); treatment100days compared with the treatment of50days,αEβ7expression was significantly higher (P=0.000), but lower than the model group expression levels at the same time point (P=0.0012).2.3The gut homing receptor αEβ7molecule expression of ileal lymphoid tissueModel group100days compared with the baseline point of model group, ileal lymphoid tissue gut homing receptor αEβ7molecule expression was significantly increased (P=0.002); medication50days compared with the baseline point of model group, αEβ7molecule expression decreased (P=0.011); medication100days in comparison with the model group100days,αEβ7molecule expression was significantly decreased (P=0.000); treatment100days compared with50days treatment,αEβ7molecule expression was not statistically significant, but it was significantly lower than the expression of the model group at the same time point (P=0.000).2.4The gut homing receptor CD62L (L-selectin)molecule expression of ileal lymphoid tissueModel group of100days compared with the baseline point of model group, intestinal lymphoid homing receptor CD62L molecule counts was on a downward trend in ileal lymphoid tissue(P>0.05), but not statistically significant; medication50days compared with the baseline point of model group, CD62L molecule counts were significantly higher (P=0.004); medication100days100days compare with the model group, CD62L molecule counts were significantly higher (P=0.002); treatment of100days compared with50days treatment, CD62L molecule counts was no statistically significant difference, but significantly higher than the model group at the same time points (P=0.002); treatment100days compared with treatment of50day, the total area of positive CD62L molecule was significantly higher (P=0.041).3. CD62L changes of AIDS monkey model in peripheral blood and CD62L changes in peripheral blood by kidney-nourishing, spleen-strengthening and detoxification therapy.CD62L plasma concentrations showed a gradual upward trend in the acute phase when Chinese rhesus macaques infected with SIV, and reach the first peak around infected eight weeks. This study found that TCM intervention could able to suppress the sudden rise of CD62L plasma concentration(P values were0.044,0.048), so CD62L plasma concentration could be maintained at a relatively stable low level state.4. The results of the HLA-DR molecules expression on peripheral blood T cells of AIDS monkey after kidney-nourishing, spleen-strengthening and detoxification therapyWith the treatment after discontinuation of2months, TCM intervention could enhance CD4percentage (P=0.04); lowered CD4+HLA-DR+percentage、 count and CD4+CD45+HLA-DR+percentage, P values were0.05,0.04,0.02, which were all statistically significant.5. The results of the αEβ7, CCR9molecular expression on peripheral blood CD4+T cells of AIDS monkey after kidney-nourishing, spleen-strengthening and detoxification therapyCD4+αEβ7+, CD4+CCR9+were suddenly increased after infected SIV one week, then decreased gradually, but there were suddenly rises after infected seven months. This study found that TCM intervention subacute AIDS stage monkey model, could inhibit CD4+CCR9+and CD4+αEβ7+suddenly increasing trend; make CD4+CCR9+, CD4+αEβ7+T cell subsets in a relatively stable state.6. The results of intestinal inflammatory cytokines-LPS changes in AIDS monkeys after kidney-nourishing, spleen-strengthening and detoxification therapyControl group:the serum concentrations of LPS were gradually increased (P<0.05) with time within six months of infection; after treatment2months, LPS serum concentrations of low dose and high dose treatment groups were lower than the control group (P<0.05); but after the withdrawal2months, there were no significant difference in the three groups; high dose group:LPS serum concentrations of administration2months was lower than before administration(P<0.05).ConclusionIn this paper, the results suggest that kidney-nourishing, spleen-strengthening and detoxification therapy on Chinese rhesus monkey model of AIDS:1. Could suppress or lower the over activation of CD4+T cells and initial CD4+T cells in peripheral blood, as well as the ileal tissue T cells.2. Could adjust the intestinal lymphatic homing homeostatic mechanisms, As shown in the data, TCM therapy inhibits the effector T lymphocytes’ intestinal lymphatic homing, but promotes the initial T lymphocytes’ intestinal lymph homing.3. Could reduce serum concentration of LPS, a kind of intestinal inflammatory factor, thereby inhibit excessive destruction of intestinal infection and inflammation caused by immune response.4. Could lower the expression of "integrin α4β7", which is a kind of HIV-1gastrointestinal receptor, then may reduce HIV-1infectivity and improve the immune function of intestinal mucosa.